ADP-Ribosylation and Regulation of Cellular Functions

ADP-核糖基化和细胞功能调节

• 批准号: 9220190
• 负责人: David Payne
• 金额: $ 20.83万
• 依托单位: Auburn University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-03-01 至 1998-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9220190&HistoricalAwards=false
• 关键词: ADP; Ribosylation; Regulation; Cellular; Functions

Currently there is reasonably convincing support for the hypothesis that reversible modification of proteins by ADP-ribosylation plays a significant role in regulation of cellular functions and signal transduction in eukaryotes. The overall objective of this research is to elucidate at the molecular level the role(s) of ADP- riblosyltransferases (ADPRTases) in signal transduction and regulation of cellular functions. To achieve this goal, it will be necessary to (1) understand how the activity of these enzymes is regulated, (2) identify the proteins which serve as physiologically relevant targets for this post-translational modification, and (3) determine how the modification of these specific proteins controls cellular functions. Progress toward these goals is hampered by the lack of immunological and molecular genetic reagents for studying the modifying enzymes and target proteins involved in ADP- ribosylation reactions. For example, no gene for an endogenous ADPRTase has yet been cloned or sequenced. Likewise, no target protein for in vivo ADP-ribosylation has been identified. One specific goal of this proposal is to generate immunological and molecular genetic reagents to use as probes in addressing some of the important questions surrounding these post-translational modifications of proteins. The parallel and complementary aims of this study are (1) generation of antibodies that specifically recognize ADPRTases and ADP-ribosylated proteins and (2) molecular analysis of these proteins and their genes. %%% Many proteins are structurally modified after synthesis by addition of various small molecules. In some cases these modifications have been shown to play a pivotal role in the mechanism of action of the protein. For example, in many cellular responses to external stimuli the mechanism of signal transduction often involves the activation of an enzyme that adds or removes a phosphate group from some protein. Another type of protein modification is the addition of a molecule, known as adenosine diphosphoribose, derived from a vitamin and a sugar, to specific proteins. This chemical reaction is carried out by an enzyme known as adenosine diphosophoribosyl- transferases (ADPRTases). The overall goal of this research is to elucidate at the molecular level the role(s) of ADP- ribosyltransferases in signal transduction and regulation of cell functions. The near term objectives are to characterize ADP ribosyltransferases and to identify and characterize the physiologically important protein substrates of the ADPRTases. First it is necessary to develop biochemical and immunological tools to proceed with these studies. The results of this research should be an important contribution to understanding of the molecular mechanisms by which cells respond to external stimuli.

目前，ADP-核糖基化对蛋白质的可逆修饰在真核生物的细胞功能和信号转导调节中发挥着重要作用，这一假设得到了相当令人信服的支持。 本研究的总体目标是在分子水平上阐明 ADP-核糖基转移酶 (ADPRTase) 在信号转导和细胞功能调节中的作用。 为了实现这一目标，有必要（1）了解这些酶的活性是如何调节的，（2）识别作为这种翻译后修饰的生理相关靶标的蛋白质，以及（3）确定修饰如何进行这些特定蛋白质控制着细胞功能。 由于缺乏用于研究 ADP-核糖基化反应中涉及的修饰酶和靶蛋白的免疫学和分子遗传学试剂，实现这些目标的进展受到阻碍。 例如，尚未克隆或测序内源 ADPRTase 的基因。 同样，尚未鉴定出体内 ADP-核糖基化的靶蛋白。 该提案的一个具体目标是产生免疫学和分子遗传试剂，用作探针来解决围绕这些蛋白质翻译后修饰的一些重要问题。 这项研究的并行和互补目标是 (1) 生成特异性识别 ADPRTase 和 ADP 核糖基化蛋白的抗体，以及 (2) 对这些蛋白及其基因进行分子分析。 %%% 许多蛋白质在合成后通过添加各种小分子进行结构修饰。 在某些情况下，这些修饰已被证明在蛋白质的作用机制中发挥着关键作用。 例如，在许多细胞对外部刺激的反应中，信号转导机制通常涉及酶的激活，该酶在某些蛋白质中添加或去除磷酸基团。 另一种类型的蛋白质修饰是将一种称为腺苷二磷酸核糖的分子添加到特定蛋白质中，该分子源自维生素和糖。 该化学反应由一种称为腺苷二磷酸核糖基转移酶 (ADPRTase) 的酶进行。 本研究的总体目标是在分子水平上阐明 ADP-核糖基转移酶在信号转导和细胞功能调节中的作用。 近期目标是表征 ADP 核糖基转移酶，并鉴定和表征 ADPRTase 的生理上重要的蛋白质底物。 首先，有必要开发生化和免疫学工具来进行这些研究。 这项研究的结果应该对理解细胞响应外部刺激的分子机制做出重要贡献。